Expression Analysis of BB0174, A Membrane Protein of Borrelia burgdorferi, the Causative Agent of Lyme Disease

Abstract

Borrelia burgdorferi (Bb), the causative agent of Lyme disease, is a spirochetal pathogen that colonizes mammalian hosts causing numerous neurological and physiological symptoms. This thesis focuses on the gene expression of BB0174, a membrane protein of Bb. Gene expression in Bb is dependent on many environmental signals following transmission from tick to host. The analysis of BB0174 shows high in silico similarities with the typical membrane associated histidine kinases of a Two-component system (TCS) for signal transduction. Thus this study will determine whether changes in temperature and pH conditions will induce changes in the levels of mRNA and protein of BB0174 in Bb. RNA will be purified from Bb growing at differing pH and temperature conditions similar to those found in the unfed tick and again from the fed tick. cDNA will then be generated and PCR analysis using specific primers for bb0174 and neighboring regions will be conducted. The BB0174 protein levels will also be evaluated after growing Bb at room temperature and shifted to 37ºC. This project will allow us to determine whether or not BB0174 is in fact one of the members of a novel TCS, and thus further our understanding of the mechanisms orchestrating gene expression in Bb.